【摘要】： 隨著電力事業(yè)的發(fā)展,極低頻磁場(chǎng)(extremely low frequency magnetic fields, ELF MF)對(duì)生物體的可能影響及其可能致癌性受到廣泛關(guān)注,國際癌癥研究組織(IARC)于2001年將其定義為人類可疑致癌源。但對(duì)ELF MF的流行病學(xué)研究結(jié)論并不一致,ELF MF與腫瘤的關(guān)系還需要體內(nèi)外實(shí)驗(yàn)證據(jù)的支持。腫瘤發(fā)生發(fā)展是一個(gè)復(fù)雜的多階段過程,其中包括浸潤和遷入周圍組織、血管新生、淋巴細(xì)胞的遷移運(yùn)輸?shù)壬砘顒?dòng),這些過程都需要細(xì)胞增殖和遷移的參與。而微絲細(xì)胞骨架的重組和循環(huán)運(yùn)動(dòng)則為其提供了動(dòng)力和物質(zhì)基礎(chǔ)。已經(jīng)發(fā)現(xiàn),在多種腫瘤細(xì)胞中,肌動(dòng)蛋白微絲被大量修飾或出現(xiàn)過表達(dá)現(xiàn)象,并發(fā)現(xiàn)與微絲功能相關(guān)的一些重要的信號(hào)分子活性或協(xié)同表達(dá)的上調(diào)等與腫瘤的發(fā)生和轉(zhuǎn)移密切相關(guān)。 本研究選擇在腫瘤發(fā)生發(fā)展中與細(xì)胞增殖、遷移、信號(hào)轉(zhuǎn)導(dǎo)均有密切關(guān)系的微絲為主要研究對(duì)象。首先采用免疫熒光法,使用羅丹明標(biāo)記的鬼筆環(huán)肽(TRITC-Phalloidin)與纖維化肌動(dòng)蛋白(F-actin)特異性結(jié)合,觀察不同強(qiáng)度和時(shí)間工頻磁場(chǎng)輻照對(duì)中國倉鼠肺成纖維細(xì)胞(Chinese Hamster Lung, CHL)細(xì)胞F-actin的影響；第二,掃描電子顯微鏡(SEM)觀察工頻磁場(chǎng)輻照對(duì)CHL細(xì)胞表面形態(tài)的影響；第三,采用劃痕實(shí)驗(yàn)和MTT法觀察與肌動(dòng)蛋白(actin)相關(guān)的細(xì)胞遷移和增殖的變化；第四,通過免疫印跡法對(duì)其可能的信號(hào)通路機(jī)制進(jìn)行初步探索。 結(jié)果顯示50 Hz不同強(qiáng)度工頻磁場(chǎng)暴露不同時(shí)間可以誘導(dǎo)CHL細(xì)胞出現(xiàn)不同程度的F-actin骨架重組和邊緣化現(xiàn)象,同時(shí)導(dǎo)致細(xì)胞周圍偽足增多,形態(tài)扁平。0.4 mT，，24 h暴露可顯著增加細(xì)胞遷移速率,但本實(shí)驗(yàn)條件下ELF MF并不能引起PI3K和磷酸化Akt蛋白表達(dá)的顯著變化。
[Abstract]:With the development of electric power industry, the possible effect of very low frequency magnetic field (extremely low frequency magnetic fields, ELF MF) on organisms and its possible carcinogenicity have been paid more and more attention. The (IARC) of the International Organization for Research on Cancer (IARC) defined it as a suspected human carcinogenic source in 2001. However, the epidemiological findings of ELF MF do not agree with the relationship between, ELF MF and tumor, and need to be supported by in vivo and in vitro experimental evidence. Tumorigenesis and development is a complex multi-stage process, including infiltration and migration into surrounding tissues, angiogenesis, lymphocyte migration and other physiological activities, all of which require the participation of cell proliferation and migration. The recombination and cyclic movement of the microfilament cytoskeleton provide the power and material basis for the microfilament cytoskeleton. It has been found that actin microfilaments have been extensively modified or overexpressed in various tumor cells. It was also found that some important signal molecules related to the function of microfilaments or the up-regulation of co-expression were closely related to the occurrence and metastasis of tumor. In this study, microfilaments, which are closely related to cell proliferation, migration and signal transduction, were selected as the main research objects. Firstly, the specific binding of Rhodamine labeled TRITC-Phalloidin to fibroactin (F-actin) was used to observe the effect of different intensity and time frequency magnetic field irradiation on (Chinese Hamster Lung, of Chinese hamster lung fibroblasts by immunofluorescence method. CHL) cell F-actin; Secondly, scanning electron microscope (SEM) was used to observe the effect of power frequency magnetic field irradiation on the surface morphology of CHL cells, third, the changes of cell migration and proliferation associated with actin (actin) were observed by scratch test and MTT method. Fourthly, the possible mechanism of signal pathway was preliminarily explored by Western blotting. The results showed that 50 Hz exposure to 50 Hz power frequency magnetic field could induce the recombination and marginalization of F-actin cytoskeleton in CHL cells in different degrees, at the same time, the pseudopodia around the cells increased and the morphology was flat 0.4 mT,. 24 h exposure significantly increased the cell migration rate, but ELF MF did not cause significant changes in the expression of PI3K and phosphorylated Akt.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R35

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